Objective To develop a whole-genome DNA microarray based on the genomic sequences of Y. pestis CO92 and 91001 and its use in comparative genomic analysis. Methods A total number of 4 005 genes of Y. pestis were amplified by PCR and printed onto glass slides in duplicate. Fluorescently labeled probes were prepared by marking genomic DNAs with random hexamers and Klenow. Labeled DNAs were hybridized with the microarrays by the method of two-fluorescence comparative hybridization. Three sets of two-fluorescence hybridizations were performed to examine the absence/presence of each gene. Results The results agreed with those derived from the in silico genomic comparison. Conclusion The results demonstrate that the microarry can be a useful tool for comparative genomic analysis of Y. pestis.

Objective To identify and compare the genome differences among live plague vaccines prepared with different strains of the bacillus. Methods The whole-genome DNA microarray of Yersinia pestis was used as a tool to perform genomic comparison among live plague vaccines prepared with 19 different strains. Results Dozens of deletions and/or increased copies of the genomic fragments were identified in the studied vaccines of different strains. Conclusion The revealed genomic differences among the vaccines from different origins account for the variability of the immunogenic and protective potency of live plague vaccines. The whole-genome DNA microarray was also provesd to be an ideal tool for the pre-evaluation of a vaccine strain.

Objective To understand the mutation characteristics of drug resistance-associated genes rpoB, katG and inhA in Mycobacterium tuberculosis (MTB) strains using gene chip method, and evaluate its clinical application value. Methods A total of 76 MTB strains were collected from Shijiazhuang area in 2013 to 2014. Gene chip was used to detect the mutations of rpoB, katG and inhA, and the L-J proportion drug susceptibility test was used as the gold standard to evaluate the overall concordance, sensitivity and specificity of gene chip. The consistency of microarray and phenotypic resistance was evaluated by Kappa test. Results Of all the 76 strains detected, 69 harbored mutations in katG/inhA. The predominant mutation site of katG was 315 codon with the mutation rate of 89.9%(62/69), and 5.8%(4/69) carried mutations at inhA-15(C→T), and 4.3%(3/69)carried combined mutations of katG 315 and inhA-15. The rpoB mutations were detected in 73 strains, of which 64.4%(47/73)carried mutations at codon 531, 15.1%(11/73)at codon 526, 12.3%(9/73)at 516 codon, 1.4%(1/73)at 513 codon, 1.4%(1/73)at 533 codon and 5.5%(4/73)had combined mutations. Compared with results from the L-J proportion method, the sensitivity, specificity and concordance rates of gene chip for RFP were 96.1%(73/76), 100%(50/50)and 97.6%(123/126). The sensitivity, specificity and concordance rates of gene chip for INH were 90.8%(69/76), 100%(50/50)and 94.4%(119/126). The sensitivity, specificity and concordance rates of gene chip for MDR-TB were 86.8%(66/76), 100%(50/50) and 92.1%(116/126). Conclusion The predominant mutation loci of MDR strains in Shijiazhuang area are katG315 and rpoB531. Gene chip is a fast and useful tool for clinical diagnosis of MDR strains.

Objective To study the effect of rimonabant, cannabinoid receptor 1(CB1) antagonist, on the expressions of CB1 andα-smooth muscle actin (α-SMA) in C57 mice with experimental hepatic fibrosis, and their mechanisms in liver fibrosis progression thereof. Methods Thirty C57 mice were randomly divided into three groups, normal control group, mod-el control group and model+rimonabant group, 10 mice for each group. The mouse model of experimental hepatic fibrosis was induced by intraperitoneal injection with 10%CCl4 for two weeks. The normal saline was delivered by gavage daily in normal control group and model control group. Rimonabant was given to mice in model+rimonabant group. Mice were sacri-ficed at the end of eight weeks. Samples of liver tissue were collected. The expressions of CB1 andα-SMA in liver tissue of mice were observed by immunohistochemical staining. The score of fibrosis stage (S) in liver tissue was also analyzed. Re-sults The positive expressions of CB1 andα-SMA and the score S were significantly higher in model control group and model+rimonabant group than those in normal control group (P<0.05). The positive expressions of CB1 andα-SMA and the score S were significantly lower in rimonabant group than those in model control group (P<0.05). There were positive corre-lations in CB1,α-SMA and S scores between normal control group, model control group and model+rimonabant group (P<0.05). Conclusion The activation of CB1 can promote the formation of liver fibrosis. The anti-fibrotic effect of rimonabant, CB1 antagonist, related with the inhibiting of the proliferation and activation of hepatic stellate cells (HSC), and the inhibit-ing of the expression of CB1.

Objective To investigate the regulation of the immune function of cytokine-induced killer cells(CIK) by dendritic cell (DC) vaccine in the patients with chronic hepatitis B(CHB) in vitro.Methods CIK cells from 30 patients with CHB were cultured in vitro,and were randomly divided into two groups,DC vaccine-treated group and the control group.After 14 days of culture,the percentages of CD3 + CD4+ T,CD3 +CD8 +T and CD3 +CD56+ T cells among CIKs were analyzed by flow cytometry.The levels of IL-12,IFN-γand IL-6 in cell culture supernatant was detected by ELISA.Results The percentages of CD3 + CD4 + T,CD3 +CD8+T and CD3+ CD56+ T cells were 18.27％,64.36％ and 20.00％ in CIKs in DC vaccine group,and 17.79％ ( P ＞ 0.05 ),54.69％ ( P ＜ 0.01 ) and 13.39％ ( P ＜ 0.01 ) in the control group,respectively.The perentage of CD3 + CD4 + T cells were similar between the two groups ( P ＞ 0.05 ),but for the perentage of CD3 + CD8 +T and C D3 + CD56 + T cells were significantly different between the two groups (t =4.130 and 5.601respectively,Ps ＜ 0.01 ).The concentrations of IL- 12,IL-4 and IFN-γin supernatant were ( 177.82 ± 130.06),(31.77 ± 9.52) and (86.99 ± 56.30) ng/L in DC vaccine-treated group respectively,which were significantly different from those of (80.83 ±50.15) ng/L (t =3.811,P ＜0.01 ),(40.33 ± 19.74) ng/L( t =2.141,P ＜0.05) and (42.07 ± 19.68)ng/L(t =4.125,P ＜0.01) in the control group,respectively.Conclusion DC vaccine could enhance the killing function of CIK cells.

Objective To investigate hepatic expressions and significances of cannabinoid receptor 1 (CB1) and cannabinoid receptor 2(CB2) in C57 mice with experimental cirrhosis.Methods Thirty C57 mice were randomly divided into three groups,i.e.normal control group,model control group and model colchicine group.Hepatic fibrosis model was prepared by intraperitoneal injection of carbon tetrachloride.The expressions of CB1 and CB2 in liver tissue of mice were observed by immunohistochemistry.The scores of inflammation grade (G) and fibrosis stage (S) were simultaneously performed.Results The scores of G and S in model control group and model colchicine group were significantly higher than those in normal control group( F =125.41,P =0.00; F =99.18,P =0.00).The scores of G and S in model control group were significantly higher than those in model colchicine group(P ＜0.01 ).The scores of CB1 and CB2 expressions in model control group and model colchicine group were significantly higher than those in normal control group ( F =29.27,P =0.00; F =36.99,P =0.00).The scores of CB1 and CB2 in model control group were significantly higher than those in model colchicine group( P ＜ 0.05 or P ＜ 0.01 ).There were significant relationships among scores of CB1,CB2,G and S in model control group and model colchicine group(Ps ＜0.05).As the scores of G and S became higher,the expressions of CB1 and CB2 gradually became more intensive.Conclusion The hepatic expressions of CB1 and CB2 in C57 mice with experimental cirrhosis increased significantly and have significant relationship with the grades of liver tissue inflammation and fibrosis.

Objective To investigate the alterations of Treg cells , Th17 cells and related cyto-kines in peripheral blood of patients during the early stage of hand-foot-mouth disease ( HFMD) .Methods Flow cytometry was performed to analyze the percentages of Treg cells ( CD4+CD25+Foxp3 T cells) and Th17 cells ( CD3+CD8-IL-17+T cells) in peripheral blood samples collected from 49 patients with severe HFMD , 26 patients with common HFMD and 30 healthy children.The levels of IL-6, IL-10, IL-17, IL-23 and TGF-β1 in serum samples were measured by ELISA .Results The percentages of Treg cells , ratios of Treg/Th17 cells, serum levels of TGF-β1 and IL-10 in patients with HFMD were significantly decreased as compared with those of control group (F=5.580, 6.205, 0.000, 0.014, respectively, P<0.05).Patients with se-vere HFMD showed a significantly increased Th17 cells (F=3.189 P<0.05) and a tendency of enhanced IL-17 expression , but no significant differences with the levels of IL-17 were observed .No significant differ-ences with the expression of IL-23 in the patients among each group were detected (P>0.05).The levels of IL-6 in serum samples from severe disease group were obviously increased as compared with those of common HFMD group and control group (F=7.318, P<0.05).Conclusion The results of this study demonstrated that the levels of Treg , Th17 cells and some related cytokines were varied in peripheral blood of patients dur-ing the early stage of HFMD .Inflammatory responses were enhanced to promote anti-virus activities by sup-pressing Treg cells and stimulating Th17 cells.

Objective To investigate expressions of surface maturation markers, secreting cytokines and the stimulat?ing effect on T lymphocytes in the dendritic cells (DC) from peripheral blood of tuberculosis (TB) patients after loading dose treatment. Methods TB patients who received initial treatment (n=68) were collected at the fifth hospital of Shijiazhuang from 2013 June to 2014 January. Base on clinical diagnosis and treatment guidelines of tuberculosis, they were divided into sputum smear-negative group (35 cases) and sputum smear-positive group(33 cases). Forty cases of healthy adult were se?lected as control group. Mononuclear cells were isolated from peripheral blood and were cultured in medium to differentiate into DCs. Expression levels of CD83 and CD86 on DCs were examined by flow cytometry. The proliferation of allogeneic mixed lymphocyte stimulated by DCs was dectected using MTT assay. Contents of IL-12, IL-10 and INF-γin the cultural supernatant of DCs and blood serum from TB patients were detected by ELISA. Results Compared with controls ,the ex?pressions of CD83 and CD86 on DCs in TB patients after loading dose treatment decreased obviously(P<0.05), and the ability to stimulate the proliferation of lymphocytes reduced evidently(P<0.05). What’s more, IL-12 level increased mark?edly(P<0.05)while IL-10 and INF-γlevels presented no significant difference among the three groups (P>0.05). Conclu?sion The expressions of maturation markers of DC cells of the peripheral blood in TB patients after initial treatment de?creased. The ability of stimulating mixed lymphocyte proliferation is also significantly reduced while secretion of IL-12 was enhanced.

Objective To observe expression and location of cannabinoid receptor 1 (CB1) in liver tissue of patients with chronic hepatitis B (CHB) ,and analyze the relationship of it with the liver fibrosis score,the serum levels of TGF-β1 and Leptin. Methods Liver biopsies were performed in 118 patients with CHB.The expression of CB1 in liver tissue was observed by immune histochemical staining, and semi-quantitative analysis was carried out to devide the CB1 score into four grades: -, +, + +, + + +. Serum levels of TGF-β1 and Leptin were determined by ABC-ELISA double-antibody sandwich method. Results The expression of CB1 in liver tissue with CHB had significant relationship with the fibrosis score. As the expression of the CB1 increased, the fibrosis score became higher ( F = 23. 369,P = 0. 000). Moreover, the expression of CB1 in liver tissue with CHB had significant relationship with the serum levels of TGF-β1 and Leptin( F values were 8. 762 and 5. 749;P values were 0. 001 and 0. 027, respectively). Conclusion CB1 may play promotive role in the process of hepatic fibrosis through regulation of TGF-β1 and Leptin.

Disease Progression ; Hepatic Stellate Cells ; Hepatitis B, Chronic ; Humans ; Liver Cirrhosis ; Receptors, Cannabinoid